The role of neurpopeptide Y in the arcuate nucleus of the hypothalamus in the regulation of energy expenditure and the involvment of its peripherally expressed receptor type-1 in the regulation of energy homeostasis in mice

下丘脑弓状核神经肽Y在能量消耗调节中的作用及其外周表达的1型受体在小鼠能量稳态调节中的作用

• 批准号: 231922378
• 负责人: Dr. Giacomo Mancini
• 金额: --
• 依托单位: Neurobiology Research Program
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/231922378?language=en
• 关键词: role; neurpopeptide; arcuate; nucleus; hypothalamus

Aim 1: Does NPY deletion in different neuron populations of the arc effect energy balance and cellular fuel selection?Body composition (whole body lean and fat mass), energy expenditure and physical activity in the conditional KO and control mice will be examined by indirect calorimetry at 9 weeks of age as well as at the end of the study at 16 weeks of age. Upon completion of the study plasma, brown adipose tissue (BAT) and white adipose tissue (WAT), liver and muscle will be dissected and the expression patterns/activity levels of various key regulators involved in mitochondrial function and thermogenesis in relevant tissues, including mitochondrial respiratory chain complexes (IV), citrate synthase, peroxisome proliferator-activated receptor ý co-activator (PGC-1a), uncoupling protein-1 (UCP-1) will be analyzed by real time PCR, Western blot analysis and/or enzyme activity assay. To verify whether differences in cellular fuel selection are present, we will analyze regulators involved in lipid oxidation such as carnitine palmitoyltransferase-1 (CPT-1), beta-hydroxyacyl CoA dehydrogenase (b-HAD) and medium-chain acyl-CoA dehydrogenase (MCAD) in several organs, such as liver, WAT, muscle. Furthermore, serum will be collected and the analysis of an extensive panel of hormones involved in the regulation of metabolism will be carried out. Does NPY deletion in different neuron populations of the arc affect energy expenditure via regulation of hypothalamic peptides?In order to see if any changes in energy expenditure can be attributed to changes in expression patterns and levels of hypothalamic peptides, brains from these mice will also be collected and mRNA expression levels of different peptides will be evaluated by using in situ hybridization. Particular emphasis will be given to the analysis of the mRNA expression levels in the Arc of POMC and CART, the opposing neurotransmitters to NPY and AGRP. Aim 2. To determine the contribution of peripheral Y1 receptors to the regulation of energy homeostasis.Does peripherally deletion of Y1 receptor effect energy balance and cellular fuel selection? By using a tamoxifen-inducuble mouse line, conditional Y1 knockout mice will be analyzed on SD conditions similarly to what described in Aim1.Does peripherally deletion of Y1 receptor protects against diet-induced obesity? In order to determine the effect of tissue specific Y1-receptor deletion in an obese context, a second set of mice will be analyzed in the same way but under high fat feeding conditions. The same parameters will be analyzed as described above. It is important to test these animals also under these conditions, as it will demonstrate which Y1 expressing tissue is most affected by high calorie and high fat intake.

目标1：在不同神经量量中的NPY缺失是否会效应能量平衡和细胞燃料选择？身体成分（全身瘦和脂肪质量），有条件的KO和对照小鼠的能量消耗和体育活动将通过在9周龄以及16周的研究结束时通过间接热量法检查。研究血浆后，将解剖棕色脂肪组织（BAT）和白色脂肪组织（WAT），肝脏和肌肉，以及与线粒体呼吸链复合物（IV）的各种关键调节剂的表达模式/活性水平，包括线粒体呼吸链复合物（IV）的各种关键调节剂，柠檬酸蒸发剂，柠檬酸蒸发液，浓氧（PGC-1A），将通过实时PCR，Western blot分析和/或酶活性测定法分析解耦蛋白-1（UCP-1）。为了验证是否存在细胞燃料选择的差异，我们将分析参与脂质氧化的调节剂，例如肉碱棕榈酰转移酶-1（CPT-1），β-羟基乙酰乙酰乙酰基CoA脱氢酶（B-HAD）和中链酰基-COA脱氢酶（MCAD），例如，在几个Organs中，例如中等链氨基-COA脱氢酶（MCAD）此外，将收集血清，并将对参与代谢调节的广泛激素进行分析。弧的不同神经量的NPY缺失是否通过调节下丘脑肽会影响能量支出？以查看能量支出的任何变化是否可以归因于下丘脑肽的表达模式的变化和下丘脑肽的水平，这些小鼠的大脑还将通过使用不同肽的MRNA表达水平来评估Sere sere insue insue hybrid sy.hybrid。特别强调了POMC和CART的mRNA表达水平的分析，POMC和CART是NPY和AGRP的相反神经递质。目的2。确定周围Y1受体对能量体内稳态调节的贡献。Y1受体效应能量平衡和细胞燃料选择的外围删除吗？通过使用他莫昔芬诱导的小鼠系，将在SD条件下分析有条件的Y1基因敲除小鼠，类似于AIM1中所述的Y1受体的外周缺失，可以预防饮食诱导的肥胖症？为了确定组织特异性Y1受体缺失在肥胖环境中的影响，将以相同的方式但在高脂肪进食条件下分析第二组小鼠。如上所述，将分析相同的参数。在这些条件下还要测试这些动物很重要，因为它将证明哪种Y1表达组织受高热量和高脂摄入量最大的影响。